高速逆流色谱分离同分异构体

Separation of Isomeric Compounds by High Speed Countercurrent Chromatography

应用高速逆流色谱法对同分异构体的分离纯化进行研究。实验结果表明,以正己烷-乙酸乙酯-水(体积比为1∶10∶10)为两相溶剂系统,上相为固定相,下相为流动相,进行二次高速逆流色谱分离,可从茶多酚中分离出g级的儿茶素同分异构体———(-)-表没食子儿茶素没食子酸酯(EGCG)和(-)-没食子儿茶素没食子酸酯(GCG),其高效液相色谱纯度均在98%以上。选择四氯化碳-氯仿-甲醇-水(体积比为7∶3∶7∶3)为溶剂系统,下相为固定相,上相为流动相,经一次高速逆流色谱即可将药物中间体溴代苯胺同分异构体进行有效的分离。

High speed countercurrent chromatography （HSCCC） was successfully used for the separation and preparation of isomeric compounds. The influence of various two-phase solvent systems on the resolution of the natural catechin isomers of （ - ）-epigallocatechin gallate （EGCG） and （ - ）-gallocatechin gallate （ GCG ） from green tea polyphenols and the synthetic isomeric compounds of para-, ortho-, meta-bromoanilline was investigated. The results indicated that the catechin isomers of （ - ）-EGCG and （ - ）-GCG can be isolated through two runs of preparative HSCCC with two-phase solvent system composed of n-hexane-ethyl acetate-water （ 1： 10：10 in volume ratio）. The upper organic phase was used as stationary phase and the lower aqueous phase as mobile phase, while the mobile phase was operated at a flow rate of 3.5 mL/min, and the apparatus rotated at 800 r/min. High performance liquid chromatographic analysis of （ - ）-EGCG and （ - ）-GCG revealed that the purity was all over 98%. The isomeric compounds of para-, ortho-, meta-bromoaniline can also be effectively isolated through a single run of preparative HSCCC with the mobile phase flow rate of 3.0 mL/min and the apparatus rotation speed of 800 r/min by using tetrachloromethane-chloroform-methanol-water （ 7 ： 3 ： 7 ： 3 in volume ratio） as two-phase solvent system, while the lower phase was selected as stationary phase and the upper phase as mobile phase.